Comparison of Saccharogenic and PHADEBAS Methods for Amylase Assay in Biological Fluids

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Abstract
Amylase estimation by dinitrosalicylic acid and chromogenic substrate (PHADEBAS) methods gave comparable results for human serum. Widely differing results were obtained with urine, saliva, and pancreatic juice. Figures for the soluble starch method were 1.98, 3.15, and 4.1 times greater than for the insoluble substrate with urine, saliva, and pancreatic juice, respectively. Moreover, the amylase separated from human serum by gel filtration was 2-2.5 times less active against the chromogenic substrate than that of whole serum, i.e., gel filtration resulted in the separation of an activator which was essential for full enzyme activity on the insoluble substrate. The nature of this activator was shown to be protein. Albumin or globulin, in equimolar concentrations, restored full enzymatic activity. For accurate analysis of a-amylase in urine, saliva, pancreatic or duodenal contents by the PHADEBAS method, the incorporation of at least 3 μmol/1 albumin is recommended.

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